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Method Validation Report
Version
1
Page
6 of 49
Document Title
Validation Report for the Determination of Thiamin HCl, Riboflavin, Niacin
(Nicotinic Acid and Nicotinamide), Pantothenic acid, Vitamin B6
(Pyridoxine, Pyridoxal and Pyridoxamine) and Biotin in Infant Formula and
Adult Nutritionals by LC-MS/MS
Table 8: Linearity of Pyridoxamine
Concentration
Area
Response 1
Area
Response 2
Area
Response 3
(ng/ml)
Standard 1
0.7
0.1111
0.1088
0.1110
Standard 2
1.41
0.2231
0.2162
0.2148
Standard 3
2.81
0.4255
0.4233
0.4334
Standard 4
4.22
0.6261
0.6297
0.6347
Standard 5
5.62
0.9056
0.8677
0.8746
Standard 6
7.03
1.1190
1.0830
1.1140
Standard 7
8.44
1.3070
1.2920
1.2870
Standard 8
9.84
1.5710
1.5480
1.4990
Standard 9
11.25
1.7530
1.7350
1.7360
Standard 10
12.65
1.9900
1.9830
1.9890
Slope
(weighting: 1/x)
0.1578
0.15515
0.1547
Y-Intercept
0.0063
0.00382
-0.000301
Correlation
Coefficient
0.99955
0.99982
0.99978
Acceptance
Criteria
Correlation Coefficient (r) ≥ 0.995
Acceptable Y/N
Y
Y
Y
Table 8: Linearity of Biotin
Concentration
Area
Response 1
Area
Response 2
Area
Response 3
(ng/ml)
Standard 1
0.71
0.0544
0.0543
0.0536
Standard 2
1.41
0.1075
0.1103
0.1095
Standard 3
2.83
0.2183
0.2145
0.2235
Standard 4
4.24
0.3064
0.3167
0.3316
Standard 5
5.66
0.4148
0.4276
0.4268
Standard 6
7.07
0.5337
0.5297
0.5231
Standard 7
8.49
0.6006
0.6337
0.6157
Standard 8
9.9
0.7429
0.7173
0.7443
Standard 9
11.32
0.8040
0.8040
0.8374
Standard 10
12.73
0.9213
0.9226
0.9188
BVIT-01 (SLV)
FOR ERP/WORKING GROUP USE ONLY
DO NOT DISTRIBUTE
Page 19